α-acyl- and α-heteroatom-substituted benzene acetamide glucokinase activators

ABSTRACT

Substituted benzene acetamide compounds and pharmaceutically acceptable salts thereof are useful as glucokinase activators.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.09/905,152, filed Jul. 13, 2001, which claims priority of U.S.Provisional Patent Application No. 60/219,872, filed Jul. 20, 2000.

BACKGROUND OF THE INVENTION

This application is relevant to U.S. Ser. No. 09/526,143, filed Mar. 15,2000 (now U.S. Pat. No. 6,320,050) and U.S. Ser. No. 09/532,506, filedMar. 21, 2000.

Glucokinase (GK) is one of four hexokinases found in mammals [Colowick,S. P., in The Enzymes, Vol. 9 (P. Boyer, ed.) Academic Press, New York,N.Y., pages 1-48, 1973]. The hexokinases catalyze the first step in themetabolism of glucose, i.e., the conversion of glucose toglucose-6-phosphate. Glucokinase has a limited cellular distribution,being found principally in pancreatic β-cells and liver parenchymalcells. In addition, GK is a rate-controlling enzyme for glucosemetabolism in these two cell types that are known to play critical rolesin whole-body glucose homeostasis [Chipkin, S. R., Kelly, K. L., andRuderman, N. B. in Joslin 's Diabetes (C. R. Khan and G. C. Wier, eds.),Lea and Febiger, Philadelphia, Pa., pages 97-115, 1994]. Theconcentration of glucose at which GK demonstrates half-maximal activityis approximately 8 mM. The other three hexokinases are saturated withglucose at much lower concentrations (<1 mM). Therefore, the flux ofglucose through the GK pathway rises as the concentration of glucose inthe blood increases from fasting (5 mM) to postprandial (≈10-15 mM)levels following a carbohydrate-containing meal [Printz, R. G.,Magnuson, M. A., and Granner, D. K. in Ann. Rev. Nutrition Vol. 13 (R.E. Olson, D. M. Bier, and D. B. McCormick, eds.), Annual Review, Inc.,Palo Alto, Calif., pages 463-496, 1993]. These findings contributed overa decade ago to the hypothesis that GK functions as a glucose sensor inβ-cells and hepatocytes (Meglasson, M. D. and Matschinsky, F. M. Amer.J. Physiol. 246, E1-E13, 1984). In recent years, studies in transgenicanimals have confirmed that GK does indeed play a critical role inwhole-body glucose homeostasis. Animals that do not express GK diewithin days of birth with severe diabetes while animals overexpressingGK have improved glucose tolerance (Grupe, A., Hultgren, B., Ryan, A. etal., Cell 83, 69-78, 1995; Ferrie, T., Riu, E., Bosch, F. et al., FASEBJ, 10, 1213-1218, 1996). An increase in glucose exposure is coupledthrough GK in β-cells to increased insulin secretion and in hepatocytesto increased glycogen deposition and perhaps decreased glucoseproduction.

The finding that type II maturity-onset diabetes of the young (MODY-2)is caused by loss of function mutations in the GK gene suggests that GKalso functions as a glucose sensor in humans (Liang, Y., Kesavan, P.,Wang, L. et al., Biochem. J. 309, 167-173, 1995). Additional evidencesupporting an important role for GK in the regulation of glucosemetabolism in humans was provided by the identification of patients thatexpress a mutant form of GK with increased enzymatic activity. Thesepatients exhibit a fasting hypoglycemia associated with aninappropriately elevated level of plasma insulin (Glaser, B., Kesavan,P., Heyman, M. et al., New England J. Med. 338, 226-230, 1998). Whilemutations of the GK gene are not found in the majority of patients withtype II diabetes, compounds that activate GK and, thereby, increase thesensitivity of the GK sensor system will still be useful in thetreatment of the hyperglycemia characteristic of all type II diabetes.Glucokinase activators will increase the flux of glucose metabolism inβ-cells and hepatocytes, which will be coupled to increased insulinsecretion. Such agents would be useful for treating type II diabetes.

SUMMARY OF THE INVENTION

This invention provides an amide selected from the group consisting of acompound of the formula:

wherein R¹ and R² are independently hydrogen, halo, cyano, nitro,loweralkylthio, perfluoro lower alkylthio, lower alkyl sulfonyl, orperfluoro-lower alkyl sulfonyl, R³ is lower alkyl having from 2 to 4carbon atoms or a 5 to 7-membered ring which is cycloalkyl,cycloalkenyl, or heterocycloalkyl having one heteroatom selected fromoxygen and sulfur, R⁴ is —C(O)NHR⁵, or is R⁶, which is an unsubstitutedor mono-substituted five- or six-membered heteroaromatic ring connectedby a ring carbon atom to the amide group shown, which five- orsix-membered heteroaromatic ring contains from 1 to 3 heteroatomsselected from sulfur, oxygen or nitrogen, with one heteroatom beingnitrogen which is adjacent to the connecting ring carbon atom; with saidmono-substituted heteroaromatic ring being monosubstituted at a positionon a ring carbon atom other than adjacent to said connecting carbon atomwith a substituent selected from the group consisting of lower alkyl,halo, nitro, cyano, —(CH₂)_(n)—OR⁹, —(CH₂)_(n)—C(O)—OR¹⁰,—(CH₂)_(n)—C(O)—NH—R¹¹, —C(O)—C(O)—OR¹², —(CH₂)_(n)—NHR¹³; n is 0, 1, 2,3 or 4; R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are independently hydrogen orlower alkyl, R⁵ is hydrogen, lower alky, lower alkenyl, hydroxy loweralkyl, halo lower alkyl, —(CH₂)_(n)—C(O)—OR⁷, —C(O)—(CH₂)_(n)—C(O)—OR⁸,X is oxygen, sulfur, sulfonyl, or carbonyl; the * indicates anasymmetric carbon atom; and its pharmaceutically acceptable salts.

Preferably, the compound of formula I is in the “R” configuration at theasymmetric carbon, shown except in the case where X is carbonyl (C═O),when the preferred enantiomer is “S”.

The compounds of formula I have been found to activate glucokinase.Glucokinase activators are useful in the treatment of type II diabetes.

DETAILED DESCRIPTION OF THE INVENTION

The subject invention will now be described in terms of its preferredembodiments. These embodiments are set forth to aid in understanding theinvention but are not to be construed as limiting.

In one embodiment, this invention provides amides of formula I,comprising compounds of formulae II and III as follows:

wherein R¹ and R² are independently hydrogen, halo, cyano, nitro, loweralkylthio, perfluoro lower alkylthio, lower alkyl sulfonyl, orperfluoro-lower alkyl sulfonyl, (preferably hydrogen, halo, lower alkylsulfonyl, or perfluoro lower alkyl sulfonyl) R³ is a 5 to 7-memberedring which is cycloalkyl, cycloalkenyl, or heterocycloalkyl having oneheteroatom selected from oxygen and sulfur, R⁵ is lower alkyl, X isoxygen, sulfur, sulfonyl or carbonyl, the * indicates an asymmetriccarbon atom and

wherein R¹ and R² are independently hydrogen, halo, cyano, nitro, loweralkylthio, perfluoro lower alkyl thio, lower alkyl sulfonyl, orperfluoro-lower alkyl sulfonyl, (preferably hydrogen, halo, lower alkylsulfonyl, or perfluoro lower alkyl sulfonyl) R³ is a 5 to 7-memberedring which is cycloalkyl, cycloalkenyl, or heterocycloalkyl having oneheteroatom selected from oxygen and sulfur, R⁶ is an unsubstituted five-or six-membered heteroaromatic ring connected by a ring carbon atom tothe amide group shown, which five- or six-membered heteroaromatic ringcontains from 1 to 3 heteroatoms selected from sulfur, oxygen ornitrogen, with one heteroatom being nitrogen which is adjacent to theconnecting ring carbon atom, X is oxygen, sulfur, sulfonyl or carbonyl,and the * indicates an asymmetric carbon atom

Preferably, the compounds of formulae II and III are in the “R”configuration at the asymmetric carbon shown except in the case where Xis carbonyl (C═O), when the preferred enantiomer is “S”. Thepharmaceutically acceptable salts of each amide of this invention arecompounds of this invention.

In preferred amides of formula II, R¹ and R² are independently halo orlower alkyl sulfonyl, R³ is a 5 to 7-membered ring which is cyclopentyl,cyclohexyl, cyclohexenyl, or heterocycloalkyl having one heteroatomselected from oxygen and sulfur (preferably oxygen) (Compound A).

In certain amides of Compound A, R⁵ is methyl, and X is oxygen. Morepreferably R¹ and R² are independently chloro or methyl sulfonyl (whichmeans R¹ and R² may each be chloro or methyl sulfonyl, or one is chlorowhile the other is methyl sulfonyl) (compound A-1). Examples of suchcompounds where R¹ and R² are chloro are

1-[cyclopentyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea,

1-[cyclohexyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea,

1-[(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea and

[1-[(3,4-dichloro-phenyl)-(tetrahydro-pyran-4-yloxy)-acetyl]-3-methyl-urea.

Examples of the amides of Compound A-1 where R¹ is chloro and R² ismethyl sulfonyl are

1-[(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetyl]-3-methyl-ureaand

1-[(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetyl]-3-methyl-urea.

In preferred amides of formula III, R¹ and R² are independently halo orlower alkyl sulfonyl, R³ is a 5 to 7-membered ring which is cyclopentyl,cyclohexyl, cyclohexenyl, or heterocycloalkyl having one heteroatomselected from oxygen and sulfur (preferably oxygen)(Compound B).Preferably R⁶ is thiazolyl or pyridinyl, and R¹ and R² are independentlychloro or methyl sulfonyl (Compound B-1).

In certain amides of Compound B-1, it is preferred that X is oxygen,especially when R¹ and R² are chloro and R⁶ is thiazolyl or pyridinyl.Examples of such compounds where R⁶ is thiazolyl are:

2-(3,4-dichloro-phenyl)-2-(tetrahydro-pyran-4-yloxy)-N-thiazol-2-yl-acetamide,

2-Cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide,

2-Cyclohexyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide, and

2-(Cyclohex-2-enyloxy)-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide.

An example of such compounds where R⁶ is pyridinyl is2-Cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-pyridin-2-yl-acetamide.

In another amide of Compound B-1 where X is oxygen, R¹ is chloro and R²is methyl sulfonyl. Examples of such compounds are:

2-(3-chloro-4-methanesulfonyl-phenyl)-2-cyclopentyloxy-N-thiazol-2-yl-acetamideand

2-(3-chloro-4-methanesulfonyl-phenyl)-2-(cyclohex-2-enyloxy-N-(4,5-dihydro-thiazol-2-yl-acetamide.

In yet another amide of Compound B-1, X is sulfur, sulfonyl or carbonyl,R¹ and R² are chloro, and R³ is cyclopentyl. Examples of such compoundsare:

3-Cyclopentyl-2-(3,4-dichloro-phenyl)-3-oxo-N-thiazol-2-yl-propionamide,

2-Cyclopentanesulfonyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamideand

2-Cyclopentylsulfanyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide.

For each compound described above, each variable which is specificallyindicated may be combined with any other variable of formula I or may becombined with any one or more specifically indicated variable.

In the compound of formula I, the * indicates the asymmetric carbon. Thecompound of formula I may be present either as a racemate or in the “R”configuration at except in the case where X is carbonyl (C═O), when thepreferred enantiomer is “S”. the asymmetric carbon shown. The “R”enantiomers are preferred, Where R³ is asymmetric an additional chiralcenter at the ring carbon connected with X is generated. At this centerthe compounds of formula I may be present as a racemate or in the “R” or“S” configuration.

As used herein, the term “halogen” and the term “halo”, unless otherwisestated, designate all four halogens, i.e. fluorine, chlorine, bromineand iodine. Preferred halogens are chlorine and bromine, most preferredis chlorine.

As used throughout this application, the term “lower alkyl” includesboth straight chain and branched chain alkyl groups having from 1 to 7carbon atoms, such as methyl, ethyl, propyl, isopropyl, preferablymethyl. As used herein, “lower alkyl sulfonyl” means a lower alkyl groupas defined above bound to the rest of the molecule through the sulfuratom in the sulfonyl group. Similarly “perfluoro-lower alkyl sulfonyl”means a perfluoro-lower alkyl group as defined above bound to the restof the molecule through the sulfur atom in the sulfonyl group.

As used herein, “lower alkyl thio” means a lower alkyl group as definedabove where a thio group is bound to the rest of the molecule. Similarly“perfluoro-lower alkyl thio” means a perfluoro-lower alkyl group asdefined above where a thio group is bound to the rest of the molecule.

As used herein, “cycloalkyl” means a saturated hydrocarbon ring havingfrom 3 to 10 carbon atoms, preferably from 5 to 7 carbon atoms.Preferred cycloalkyls are cyclopentyl and cyclohexyl. As used herein,“cycloalkenyl” means a cycloalkyl ring having from 3 to 10, andpreferably from 5 to 7 carbon atoms, where one of the bonds between thering carbons is unsaturated. As used herein, “heterocycloalkyl” means asaturated hydrocarbon ring having from 3 to 10 carbon atoms, preferablyfrom 5 to 7 carbon atoms, and having a heteroatom which may be oxygen orsulfur. It is preferred to have a single heteroatom, preferably oxygen.

As used herein, the term “lower alkenyl” denotes an alkylene grouphaving from 2 to 6 carbon atoms with a double bond located between anytwo adjacent carbons of the group. Preferred lower alkenyl groups areallyl and crotyl.

The variable X may be an oxygen or sulfur (i.e. —O— or —S—) or sulfonylor carbonyl (i.e. SO₂ or C═O).

The heteroaromatic ring can be an unsubstituted or mono-substitutedfive- or six-membered heteroaromatic ring having from 1 to 3 heteroatomsselected from the group consisting of oxygen, nitrogen, or sulfur andconnected by a ring carbon to the amide group shown. The heteroaromaticring has at least one nitrogen atom adjacent to the connecting ringcarbon atom and if present, the other heteroatoms can be sulfur, oxygenor nitrogen. Certain preferred rings contain a nitrogen atom adjacent tothe connecting ring carbon and a second heteroatom adjacent to theconnecting ring carbon or adjacent to said first heteroatom. Theheteroaromatic rings are connected via a ring carbon atom to the amidegroup. The ring carbon atom of the heteroaromatic ring which isconnected via the amide linkage cannot contain any substituent.Heteroaromatic rings include, for example, pyrazinyl, pyridazinyl,isoxazolyl, isothiazolyl, pyrazolyl, pyridinyl, pyrimidinyl,thiadiazolyl (preferably 1,3,4-, 1,2,3-, 1,2,4-), triazinyl (preferably1,3,5-, 1,2,4-), thiazolyl; oxazolyl, and imidazolyl. Preferred ringsare thiazolyl for example 4 or 5-halothiazolyl, 4 or 5 lower alkylthiazolyl, pyridinyl, and pyrimidinyl, for example 2-lower alkylpyrimidinyl. Most preferred are thiazolyl or pyridinyl.

Preferable compounds in accordance with the present invention arecompounds of above formula I, wherein R⁵ is lower alkyl, preferablymethyl. In one embodiment, preferable heteroaromaric ring R⁶ isthiazolyl; in another embodiment, preferable heteroaromatic ring R⁶ ispyridinyl. In one embodiment, preferable R¹ and R² are independentlyhalo (preferably chloro) or lower alkyl sulfonyl (preferably methylsulfonyl); in another embodiment, R¹ and R² are chloro; in still anotherembodiment, R¹ is chloro and R² is methyl sulfonyl. Preferable residueR³ is cyclopentyl, cyclohexyl, cyclohexenyl, with cyclopentyl beingpreferred, or a six-membered heterocycloalkyl having one heteroatomselected from oxygen and sulfur, with oxygen being preferred. In oneembodiment, X is oxygen; in another embodiment, X is sulfur, sulfonyl orcarbonyl.

Most preferable compounds in accordance with the present invention are:

1-[cyclopentyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea,

1-[cyclohexyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea,

1-[(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea,

[1-[(3,4-dichloro-phenyl)-(tetrahydro-pyran-4-yloxy)-acetyl]-3-methyl-urea,

1-[(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetyl]-3-methyl-urea,

1-[(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetyl]-3-methyl-urea,

2-(3,4-dichloro-phenyl)-2-(tetraydro-pyran-4-yloxy)-N-thiazol-2-yl-acetamide,

2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide,

2-cyclohexyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide,

2-(cyclohex-2-enyloxy)-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide,

2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-pyridin-2-yl-acetamide,

2-(3-chloro-4-methanesulfonyl-phenyl)-2-cyclopentyloxy-N-thiazol-2-yl-acetamide,

2-(3-chloro-4-methanesulfonyl-phenyl)-2-(cyclohex-2-enyloxy-N-(4,5-dihydro-thiazol-2-yl-acetamide,

3-cyclopentyl-2-(3,4-dichloro-phenyl)-3-oxo-N-thiazol-2-yl-propionaimide,

2-cyclopentanesulfonyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamideand

2-cyclopentylsulfanyl-2-(3,4-dichloro-phenyl)-N-thiazol-0.2-yl-acetamide.

The term “pharmaceutically acceptable salts” as used herein include anysalt with both inorganic or organic pharmaceutically acceptable acidssuch as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid,phosphoric acid, citric acid, formic acid, maleic acid, acetic acid,succinic acid, tartaric acid, methanesulfonic acid, para-toluenesulfonic acid and the like. The term “pharmaceutically acceptable salts”also includes any pharmaceutically acceptable base salt such as aminesalts, trialkyl amine salts and the like. Such salts can be formed quitereadily by those skilled in the art using standard techniques. Thisinvention includes the pharmaceutically acceptable salt of each compoundof formula I.

The compound of formula I can be prepared by the following ReactionSchemes which follow.

During the course of the reactions, the various functional groups suchas the free carboxylic acid or hydroxy groups will be protected viaconventional hydrolyzable ester or ether protecting groups. As usedherein the term “hydrolyzable ester or ether protecting groups”designates any ester or ether conventionally used for protectingcarboxylic acids or alcohols which can be hydrolyzed to yield therespective hydroxyl or carboxyl group. Exemplary ester groups useful forthe protection of a hydroxyl group are those in which the acyl moietiesare derived from a lower alkanoic, aryl lower alkanoic, or lower alkanedicarboxcyclic acid. Among the activated acids which can be utilized toform such groups are acid anhydrides, acid halides, preferably acidchlorides or acid bromides derived from aryl or lower alkanoic acids.Example of anhydrides are anhydrides derived from monocarboxylic acidsuch as acetic anhydride, benzoic acid anhydride, and lower alkanedicarboxcyclic acid anhydrides, e.g. succinic anhydride. Suitable etherprotecting groups for alcohols are, for example, the tetrahydropyranylethers such as 4-methoxy-5,6-dihydroxy-2H-pyranyl ethers. Others arearoyl substituted methyl ethers such as benzyl or trityl ethers orα-lower alkoxy lower alkyl ethers, for example, methoxymethyl or allylicethers or alkyl silylethers such as trimethylsilylether.

Exemplary ester groups useful for the protection of carboxylic acidgroups are those derived from lower alkanols or substituted orunsubstituted benzyl alcohols. The choice of ester functions used iswell known to those of ordinary skill in the art of organic chemistry.For example, the ester functions most readily cleaved under basichydrolysis are those derived from lower primary alcohols such as methyl,ethyl, and the like. Ester functions derived from secondary or tertiaryalcohols are more readily cleaved under acidic conditions, for exampletertiary butyl or diphenylmethyl esters. Benzyl esters are particularlyuseful for the protection of carboxylic acid functions in compounds thatare stable to the hydrogenolytic conditions that can be used to removethe protecting group.

The term “amino protecting group” designates any conventional aminoprotecting group which can be cleaved to yield the free amino group. Thepreferred protecting groups are the conventional amino protecting groupssuch as those utilized in peptide synthesis, particularly thecarbamates. Particularly preferred amino protecting groups in this classare t-butoxycarbonyl (BOC), carbobenzyloxy (CBZ), and9-fluorenylmethoxy-carbonyl (FMOC) moieties. Each of these protectinggroups is readily removed under reaction conditions that do not affectthe others. For example FMOC and CBZ protecting groups are stable to theacidic conditions used to remove BOC groups and other acid labilemoieties. CBZ groups can be removed by hydrogenolysis in the presence ofFMOC and BOC protecting groups, while the FMOC moiety is particularlylabile in the presence of secondary cyclic amines, conditions underwhich BOC and CBZ groups are unaffected.

Reaction Scheme II outlines the preparation of the phenylpyruvic acidester of formula 6, from which compounds of formula I where X=O, S, orSO₂ can be prepared. The compounds of formula 6 are accessible from thecorresponding phenyl acetic acids of structure 3 or substituted benzenesof structure 1 as outlined in Reaction Scheme II (see for example,Anderson, J. C. and Smith, S.C. Syn. Lett., 1990, 107; Davis, F. A.,Haque, M. S., et al., J. Org. Chem, 1986, 51, 2402; Tanaka, M.;Kobayashi, T. and Sakakura, T.; Angew. Chem. Int. Ed. Engl, 1984, 23,518; Murahashi, S. and Naota, T., Synthesis, 1993, 433). The method toprepare the pyruvates of structure 6 via the α-hydroxy phenylaceticacids of structure 7 may be considered a general procedure regardless ofthe nature of the substituents R¹ and R², with the proviso that thesesubstituents are protected during the process with suitable protectinggroups if required. The alternative procedure, the preparation of thepyruvates of structure 6 by an electrophilic substitution reaction onthe substituted benzenes of structure 10 under Friedel-Crafts, is usefulfor certain selected R¹ and R² which can be identified by the skilledchemist.

In the compounds of formula 3 wherein one of R¹ and R² is nitro, chloro,bromo, or iodo and the other is hydrogen, either the carboxylic acids 3or their lower alkyl esters 4 (R^(a)=lower alkyl) are commerciallyavailable. In those cases where the available starting acids of formula3 or the commercially available potential progenitors 1,3, or 5 do notcarry the desired substituents, that is, R¹ and R² do not fall withinthe scope of the all definitions listed herein for R¹ and R², thesubstituents of the available starting materials can be manipulated byany of the commonly known methods to interconvert aromatic substituentsto ultimately lead to the desired substitution pattern in thephenylpyruvates of structure 6 i.e., for all definitions of R¹ and R².In cases where only the carboxylic acids of structure 3 are available,they can be converted to the corresponding esters 4 of lower alkylalcohols using any conventional esterification methods. All thesubstituent interconversion reactions discussed hereto forward arecarried out on lower alkyl esters of the compounds of formula 4.

The amino substituted compounds of formula 4 which in turn can beobtained from the corresponding NO₂ compound which can be diazotized toyield the corresponding diazonium compound, which in situ can be reactedwith the desired lower alkyl thiol, perfluoro-lower alkyl thiol (see forexample, Baleja, J. D. Synth. Comm. 1984, 14, 215; Giam, C. S.;Kikukawa, K., J. Chem. Soc, Chem. Comm. 1980, 756; Kau, D.; Krushniski,J. H.; Robertson, D. W, J. Labelled Compd Rad. 1985, 22, 1045; Oade, S.;Shinhama, K.; Kim, Y. H., Bull Chem Soc. Jpn. 1980, 53, 2023; Baker, B.R.; et al, J. Org. Chem. 1952, 17, 164), or alkaline earth metalcyanide, to yield corresponding compounds of formula 4, where one of thesubstituents is lower alkyl thio, perfluoro-lower alkyl thio, or cyano,and the other is hydrogen. If desired, the lower alkyl thio orperfluoro-lower alkyl thio compounds can then be converted to thecorresponding lower alkyl sulfonyl or perfluoro-lower alkyl sulfonylsubstituted compounds of formula 4. Any conventional method of oxidizingalkyl thio substituents to sulfones can be utilized to effect thisconversion.

In the compounds of formula 3 wherein both of R¹ and R² are chloro orfluoro, the carboxylic acids 4 or the corresponding lower alkyl estersof structure 4 are commercially available. In cases where only thecarboxylic acids are available, they can be converted to thecorresponding esters of lower alkyl alcohols using any conventionalesterification method. As shown in Reaction Scheme II, to produce thecompound of formula 3 where both R¹ and R² are nitro, 3,4-dinitrotoluene(R¹═R²═NO₂) can be used as starting material. This can be converted tothe corresponding 3,4-dinitrobenzoic acid 2. Any conventional method ofconverting an aryl methyl group to the corresponding benzoic acid can beutilized to effect this conversion (see for example, Clark, R. D.;Muchowski, J. M.; Fisher, L. E.; Flippin, L. A.; Repke, D. B.; Souchet,M, Synthesis, 1991, 871). The benzoic acids of structure 2 can behomologated to the corresponding phenyl acetic acids of structure 3 bythe well-known Arndt Eistert method.

The compounds of formula 4b where both R¹ and R² substituents are aminocan be obtained from the corresponding di-nitro compound of formula 4a,described above. Any conventional method of reducing a nitro group to anamine can be utilized to effect this conversion. The compound of formula4b where both R¹ and R² are amine groups can be used to prepare thecorresponding compound of formula 4d where both R¹ and R² are iodo,bromo, chloro, or fluoro via the diazotization reaction intermediate 4cdescribed before. Any conventional method of converting amino group toan iodo or bromo group (see for example, Lucas, H. J.; Kennedy, E. R.Org. Synth. Coll. Vol, II 1943, 351) can be utilized to effect thisconversion.

If it is desired to produce compounds of formula 4e,f, where both R¹ andR² are lower alkyl thio or perfluoro-lower alkyl thio groups, thecompound of formula 4b where R¹ and R² are amino can be used as startingmaterial. Any conventional method of converting an aryl amino group toaryl thioalkyl group can be utilized to effect this conversion. If it isdesired to produce compounds of formula 4g,h where R¹ and R² are loweralkyl sulfonyl or perfluoro-lower alkyl sulfonyl, the correspondingcompounds of formula 4e,f where R¹ and R² are lower alkyl thio orperfluoro-lower alkyl thio can be used as starting material. Anyconventional method of oxidizing alkyl thio substituents to sulfones canbe utilized to effect this conversion.

If it is desired to produce compounds of formula 4i, where both R¹ andR² are cyano groups, the compound of formula 4b can be used as startingmaterial. Any conventional method used to convert an amino group tocyano group can be utilized to effect this conversion.

The carboxylic acids of formula 3 where one of R¹ and R² is nitro andthe other is halo (for example chloro) are known from the literature(see for 4-chloro-3-nitrophenyl acetic acid, Tadayuki, S.; Hiroki, M.;Shinji, U.; Mitsuhiro, S. Japanese patent, JP 71-99504, ChemicalAbstracts 80:59716; see for 4-nitro-3-chlorophenyl acetic acid, Zhu, J.;Beugelmans, R.; Bourdet, S.; Chastanet, J.; Rousssi, G. J. Org. Chem.1995, 60, 6389; Beugelmans, R.; Bourdet, S.; Zhu, J. Tetrahedron Lett.1995, 36, 1279). These carboxylic acids can be converted to thecorresponding lower alkyl esters 4m,n using any conventionalesterification methods. Thus, if it is desired to produce the compoundof formula 4 where one of R¹ and R² is nitro and the other is loweralkyl thio (4o,p) or perfluoro-lower alkyl thio (4q,r), thecorresponding compound where one of R¹ and R² is nitro and the other ischloro can be used as starting material. In this reaction, anyconventional method of nucleophilic displacement of aromatic chlorinegroup with a lower alkyl thiol can be used (see for example, Singh, P.;Batra, M. S.; Singh, H, J. Chem. Res. —S 1985 (6), S204; Ono, M.;Nakamura, Y.; Sata, S.; Itoh, I, Chem. Lett, 1988, 1393; Wohrle, D.;Eskes, M.; Shigehara, K.; Yamada, A, Synthesis, 1993, 194; Sutter, M.;Kunz, W, US patent, U.S. Pat. No. 5,169,951). Once the compounds offormula 4 where one of R¹ and R² is nitro and the other is lower alkylthio or perfluoro-lower alkyl thio are available, they can be convertedto the corresponding compounds of formula 4 wherein one of R¹ and R² isnitro and the other is lower alkyl sulfonyl (4s,t) or perfluoro-loweralkyl sulfonyl (4u,v) using conventional oxidation procedures.

If it is desired to produce compounds of formula 4aa-ad where one of R¹and R² is lower alkyl thio and the other is perfluoro-lower alkyl thio,the corresponding compound where one of R¹ and R² is amino and the otheris lower alkylthio (4w,x) or perfluoro-lower alkylthio (4y,z) can beused as starting materials. Any conventional method of diazotizing anaromatic amino group and reacting it in situ with the desired loweralkyl thiol or perfluoroalkyl thiol can be utilized to effect thisconversion.

If it is desired to produce compounds of formula 4 where one of R¹ andR² is lower alkyl sulfonyl and the other is perfluoro-lower alkylsulfonyl, (4ae-4ah) the corresponding compounds (4aa-ad) where one of R¹and R² is lower alkyl thio and the other is perfluoro-lower alkyl thio,can be used as starting materials. Any conventional method of oxidizingan aromatic thio ether group to the corresponding sulfone group can beutilized to effect this conversion.

If it is desired to produce compounds of formula 4 where one of R¹ andR² is halo and the other is lower alkyl thio 4ai,aj) or perfluoro-loweralkyl thio (4ak,al), the corresponding compounds where one of R¹ and R²is amino and the other is lower alkyl thio (4w,x) or perfluoro-loweralkyl thio (4y,z) can be used as starting materials. Any conventionalmethod of diazotizing an aromatic amino group and conversion of it insitu to an aromatic halide can be utilized to effect this conversion.

If it is desired to produce compounds of formula 4 where one of R¹ andR² is cyano, and the other is halo, (4aq, 4ar), the correspondingcompounds of formula (4as, 4at) where one of R¹ and R² is nitro, and theother is amino can be used as starting materials. This transformationcan be achieved via conversion of amino group of compounds of formula(4as, 4at) to corresponding halo compounds (4au, 4av), which in turnfurther can be transformed to the compounds of formula (4aq, 4ar).

If it is desired to produce compounds of formula 4 where one of R¹ andR² is cyano, and the other is lower alkylthio or lower perfluoro loweralkylthio (4ba-4be), the corresponding compounds of formula 4as, 4at canbe used as starting material. Any conventional means of converting anamino group to a thioalkyl group can be used to affect this conversion.

If it is desired to produce compounds of formula 4 where one of R¹ andR² is cyano and the other is lower alkylsulfonyl orperfluoro-loweralkylsulfonyl (4bf-4bi), the corresponding compounds offormula (4ba-4be) can be used as starting material. Any conventionalmeans of converting a thio ether to the corresponding sulfone can beused to affect this conversion.

If it is desired to produce compounds of formula 4 where one of R¹ andR² is halo and the other is lower alkyl sulfonyl or perfluoro-loweralkyl sulfonyl, (4am-4ap) the corresponding compounds where one of R¹and R² is halo and the other is lower alkyl thio (4ai,aj) orperfluoro-lower alkyl thio (4ak,al) can be used as starting materials.Any conventional method of oxidizing an aromatic thio ether to thecorresponding sulfone can be utilized to effect this conversion.

In cases where one or both of R¹ or R² is an amino group in compounds ofstructure 6, the amino groups are protected with a conventional aminoprotecting group, before further transformations are carried out.

Preparation of compounds of formula I where X is O or S is outlined inReaction Scheme I. The pyruvate esters of formula 6 are transformed tothe corresponding aryl sulfonyl hydrazones of formula 9 by reacting thepyruvate esters with the appropriate sulfonylhydrazide derivative. Thisreaction is conveniently carried out by conventional aryl sulfonylhydrazide condensation reaction conditions, for example by refluxing asolution of the pyruvate ester 6 and p-toluenesulfonyl hydrazide in aninert solvent, preferably an aromatic hydrocarbon, for example benzeneor toluene, preferably toluene. The reaction may be performed in anapparatus designed such that the refluxing solvent, which contains theazeotroped reaction byproduct, water, to pass though a water removingagent, such as molecular sieves, before returning to the reaction flask.In this manner, the hydrazone forming reaction may be accelerated anddriven to completion. The p-toluenesulfonylhydrazones of formula 9, canthen be treated with an tertiary amine base in a polyhalogenated organicsolvent, for example triethylamine or diisopropylethylamine, preferablytriethylamine in a chlorinated hydrocarbon solvent, for exampledichloromethane, to give the corresponding diazo esters of formula II.This conversion is normally carried out at a temperature of between zerodegrees and 40° C., preferably at the ambient temperature.

Compounds of structure 12 where X is O may be prepared by reacting thediazo ester of formula II with the appropriate cycloalkyl, cycloalkenylor non-aromatic heterocyclic alcohol in the presence of catalytic amountof rhodium (II) acetate. The reaction is conveniently carried in aninert solvent, preferably dichloromethane at a temperature of betweenzero degrees and 40° C., preferably at room temperature.

In a like manner, compounds of structure 12, where X is S, may beprepared by reacting the diazo ester of formula II with the appropriatecycloalkyl, cycloalkenyl or non-aromatic heterocyclic mercaptan in thepresence of catalytic amount of rhodium (II) acetate. The reaction isconveniently carried in an inert solvent, preferably dichloromethane ata temperature of between zero degrees and the reflux temperature of themixture, preferably at the reflux temperature.

Preparation of compounds of formula I where X is C(O) is outlined inReaction Scheme I. More specifically, two related methods are utilizedto prepare compounds of structure III, as shown in Reaction Scheme III,where X is C(O). In the first method, the phenylacetic acids ofstructure 3 are first converted to the corresponding ester 4 by any ofthe methods well known to those of normal competence in the field oforganic chemistry. As an example, an acid of structure 3 in an inertsolvent, for example methanol or diethyl ether or tetrahydrofuran or amixture thereof, may be treated with an excess of an ethereal solutionof diazomethane, or treatment of acid 3 with methanol in the presence ofa catalytic amount of sulfuric acid.

The thus formed ester of structure 4 may be deprotonated by with anon-nucleophilic strong base, for example lithium diisopropylamide orlithium bis(trimethylsilyl)amide, in an inert solvent, for examplediethyl ether or tetrahydrofuran, preferably tetrahydrofuran. Thedeprotonation reaction may be conveniently carried out in an inertatmosphere under ardhydrous conditions at a temperature of from −50° C.to −100° C., preferably at −78° C. The lithiated species formed in thismanner, may be reacted in situ with a cycloalkyl or cycloalkenyl acidchloride of structure 19 while the reaction temperature may bemaintained at a temperature of from −50° C. to −100° C., preferably at−78° C. to give the compound of structure 12, where X=C(O).

Cleavage of the alkali-labile ester moiety in compounds of structure 12(R^(a)=unbranched lower alkyl) may be carried out in accordance withknown procedures. For example, the esters of structure 12, are treatedwith an alkali metal hydroxide, for example potassium hydroxide, sodiumhydroxide or lithium hydroxide, preferably potassium hydroxide in aninert solvent system, for example a mixture of ethanol and water. Thesaponification reaction may be generally performed at a temperature offrom zero degrees to the reflux temperature of the mixture, preferablyat room temperature, to furnish the acids of structure 14.

The coupling of carboxylic acids of structure 14 with the amines R⁶—NH₂(13) to give the amides of structure III can be performed by usingmethods well known to one of ordinary skill in the art. For example, thereaction may be conveniently carried out by treating the carboxylic acidof structure 14 with the amine 13 in the presence of a tertiary aminebase, for example triethylamine or diethylisopropylamine and a couplingagent such as O—(1H-benzotriazo-1-yl)-1,1,3,3,-tetramethyluroniumhexafluorophosphate (HBTU) orbenzotriazol-1-yloxy(dimethylamino)phosphonium hexafluorophosphate(BOP). The reaction may be carried out in an inert solvent, such as achlorinated hydrocarbon (e.g., dichloromethane) or N,N-dimethylformamideat a temperature between zero degrees and about room temperature,preferably at about room temperature, optionally in the presence of asubstance that accelerates the rate of reaction, for example1-hydroxybenzotriazole.

Alternatively, to prepare the amides of structure III, as shown inscheme III, the carboxylic acids of structure 3 can be activated throughconversion to a mixed anhydride, which may be in turn reacted with theamine 13 in the presence of a catalyst to afford the amides of structure18, or by using standard peptide coupling reagents such as HBTU.Subsequently the amide of structure 18 may be deprotonated by with anon-nucleophilic strong base, for example lithium diisopropylamide orlithium bis(trimethylsilyl)amide, in an inert solvent, for examplediethyl ether or tetrahydrofuran, preferably tetrahydrofuran. Thedeprotonation reaction may be conveniently carried out in an inertatmosphere under anhydrous conditions at a temperature of from −50° C.to −100° C., preferably at −78° C. The thus formed lithiatedintermediate, may be reacted in situ with a cycloalkyl or cycloalkenylacid chloride of structure 19 while the reaction temperature may bemaintained at a temperature of from −50° C. to −100° C., preferably at−78° C. to give the compound of structure III, where X═C(O).

To produce the primary amides of structure 15, the carboxylic acids ofstructure 14 are converted to an activated species, preferably an acidchloride which in turn may be reacted with a protected form of ammonia,hexamethyldisilazane, to give after hydrolytic removal if thetrimethylsilyl groups in situ, the primary amides. The carboxylic acidsof structure 14 are transformed into the corresponding acid chlorides ontreatment with oxalyl chloride in an inert solvent, such as achlorinated hydrocarbon (e.g., dichloromethane) or an aromatichydrocarbon such as benzene. The reaction may be carried out in thepresence of a catalytic amount of N,N-dimethylformamide at a temperatureof between zero degrees and about room temperature, preferably at aboutzero degrees. The subsequent reaction of the intermediate acid chloridewith an excess of 1,1,1,3,3,3-hexamethyldisilazane may be carried out insitu at a temperature between zero degrees and about room temperature,preferably at about room temperature. Treatment of the formedbis(trimethylsilyl)amide with a large excess of methanol containing 5%sulfuric acid at room temperature provides the desilylated primary amideof structure 15.

The ureas of structure II are produced by three methods:

(a) reaction of the acid chlorides derived as described above from thecarboxylic acids of structure 14 with a monosubstituted urea 16

(b) by reaction of the primary amide of structure 15 with and isocyanateof structure 17

(c) by reaction of esters of formula 12 (R^(a)+lower alkyl) with amonosubstituted urea (16) in the presence of an alkali metal alkoxide.

In the first mentioned procedure, the acid chloride, derived from thecarboxylic acid of structure 14 on treatment with oxalyl chloride is asdescribed above except the reaction may be run in fluorobenzene, may bereacted in situ with urea or a monosubstituted urea (16). The reactionmay be carried out at a temperature between 50° C. and about the refluxtemperature of the mixture, preferably at about 70° C. to yield theureas of structure II. In the alternative scheme, the primary amide ofstructure 15 may be reacted with an isocyanate of structure 17, in aninert solvent such as an aromatic hydrocarbon, preferably toluene. Thereaction may be normally carried out at a temperature between 50° C. andabout the reflux temperature of the mixture, preferably at the refluxtemperature to yield the ureas of structure II.

For compounds of formula I where X is S, the thioethers of structure IIand III (X═S) may be converted to the sulfones of structure I (X═SO₂) byusing methods well known to one of ordinary skill in the field oforganic chemistry. For example, the transformation may be achieved byusing a two-step procedure. In the first step, treatment of the thioethers of structures II and III (X═S) with an oxidizing agent,preferably sodium periodate in aqueous methanol furnished theintermediate sulfoxides of structure II and III (X═SO). The reaction maybe conveniently carried out at a temperature of between zero degrees andabout room temperature, preferably at about room temperature. In thesecond step, treatment of the intermediate sulfoxides II and III (X═SO)with an oxidizing agent, preferably potassium permanganate in aqueousmethanol furnished the sulfones of structure I (X═SO₂). The reaction maybe conveniently carried out at a temperature of between zero degrees andabout room temperature, preferably at about room temperature.

The compound of formula I has an asymmetric carbon atom through whichthe group XR³ and the acid amide substituents are connected. Inaccordance with this invention, the preferred stereoconfiguration ofthis group is R, except in cases where X is carbonyl, where thepreferred enantiomer is “S”. In cases wherein R³ is asymmetric (e.g.cycloalkene), an additional chiral center at the ring carbon connectingwith atom ‘X’ is generated. At this center, racemic compounds andcompounds corresponding to both R and S configuration are part of thisinvention.

If it is desired to produce the R or the S isomer of the compound offormula I, this compound can be separated into these isomers by anyconventional chemical means. Among the preferred chemical means is toreact the compound of formula 14 (same as 14 above) with an opticallyactive base. Any conventional optically active base can be utilized tocarry out this resolution. Among the preferred optically active basesare the optically active amine bases such as alpha-methylbenzylamine,quinine, dehydroabietylamine and alpha-methylnaphthylamine. Any of theconventional techniques utilized in resolving organic acids withoptically active organic amine bases can be utilized in carrying outthis reaction.

In the resolution step, the compound of formula 14 is reacted with theoptically active base in an inert organic solvent medium to producesalts of the optically active amine with both the R and S isomers of thecompound of formula 14. In the formation of these salts, temperaturesand pressure are not critical and the salt formation can take place atroom temperature and atmospheric pressure. The R and S salts can beseparated by any conventional method such as fractional crystallization.After crystallization, each of the salts can be converted to therespective compounds of formula 14 in the R and S configuration byhydrolysis with an acid. Among the preferred acids are dilute aqueousacids, i.e., from about 0.001N to 2N aqueous acids, such as aqueoussulfuric or aqueous hydrochloric acid. The configuration of formula 14which is produced by this method of resolution is carried out throughoutthe entire reaction scheme to produce the desired R or S isomer offormula I.

The separation of R and S isomers can also be achieved using anenzymatic ester hydrolysis of any lower alkyl esters corresponding tothe compound of the formula 14 (see for example, Ahmar, M.; Girard, C.;Bloch, R, Tetrahedron Lett, 1989, 7053), which results in the formationof corresponding chiral acid and chiral ester. The ester and the acidcan be separated by any conventional method of separating an acid froman ester. The preferred method of resolution of racemates of thecompounds of the formula 14 is via the formation of correspondingdiastereomeric esters or amides. These diastereomeric esters or amidescan be prepared by coupling the carboxylic acids of the formula 14 witha chiral alcohol, or a chiral amine. This reaction can be carried outusing any conventional method of coupling a carboxylic acid with analcohol or an amine. The corresponding diastereomers of compounds of theformula 14 can then be separated using any conventional separationmethods. The resulting pure diastereomeric esters or amides can then behydrolyzed to yield the corresponding pure R or S isomers. Thehydrolysis reaction can be carried out using any conventional method tohydrolyze an ester or an amide without racemization.

On the basis of their capability of activating glucokinase, thecompounds of above formula I can be used as medicaments for thetreatment of type II diabetes. Therefore, as mentioned earlier,medicaments containing a compound of formula I are also an object of thepresent invention, as is a process for the manufacture of suchmedicaments, which process comprises bringing one or more compounds offormula I and, if desired, one or more other therapeutically valuablesubstances into a galenical administration form, e.g. by combining acompound of formula I with a pharmaceutically acceptable carrier and/oradjuvant.

The pharmaceutical compositions may be administered orally, for examplein the form of tablets, coated tablets, dragees, hard or soft gelatinecapsules, solutions, emulsions or suspensions. Administration can alsobe carried out rectally, for example using suppositories; locally orpercutaneously, for example using ointments, creams, gels or solutions;or parenterally, e.g. intravenously, intramuscularly, subcutaneously,intrathecally or transdermally, using for example injectable solutions.Furthermore, administration can be carried out sublingually or as anaerosol, for example in the form of a spray. For the preparation oftablets, coated tablets, dragees or hard gelatine capsules the compoundsof the present invention may be admixed with pharmaceutically inert,inorganic or organic excipients. Examples of suitable excipients fortablets, dragees or hard gelatine capsules include lactose, maize starchor derivatives thereof, talc or stearic acid or salts thereof. Suitableexcipients for use with soft gelatine capsules include for examplevegetable oils, waxes, fats, semi-solid or liquid polyols etc.;according to the nature of the active ingredients it may however be thecase that no excipient is needed at all for soft gelatine capsules. Forthe preparation of solutions and syrups, excipients that may be usedinclude for example water, polyols, saccharose, invert sugar andglucose. For injectable solutions, excipients that may be used includefor example water, alcohols, polyols, glycerine, and vegetable oils. Forsuppositories, and local or percutaneous application, excipients thatmay be used include for example natural or hardened oils, waxes, fatsand semi-solid or liquid polyols. The pharmaceutical compositions mayalso contain preserving agents, solubilising agents, stabilising agents,wetting agents, emulsifiers, sweeteners, colorants, odorants, salts forthe variation of osmotic pressure, buffers, coating agents orantioxidants. As mentioned earlier, they may also contain othertherapeutically valuable agents. It is a prerequisite that all adjuvantsused in the manufacture of the preparations are non-toxic.

Preferred forms of use are intravenous, intramuscular or oraladministration, most preferred is oral administration. The dosages inwhich the compounds of formula (a) are administered in effective amountsdepend on the nature of the specific active ingredient, the age and therequirements of the patient and the mode of application. In general,dosages of about 1-100 mg/kg body weight per day come intoconsideration.

All of the compounds described in the following syntheses activatedglucokinase in vitro in accordance with the assay described in theBiological Activity Example.

This invention will be better understood from the following examples,which are for purposes of illustration and are not intended to limit theinvention defined in the claims that follow thereafter.

EXAMPLES Example 1 Preparation ofrac-2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide

A solution of aluminum chloride (19.96 g 149.6 mmol) in dichloromethane(85 mL) was cooled to 0° C. and then methyl oxalyl chloride (6.6 mL71.43 mmol) was slowly added and the mixture was stirred at 0-5° C. for1 h. 1,2-dichlorobenzene (7.7 mL, 68.03 mmol) was added, while thereaction temperature was maintained below 5° C. throughout the addition.After the mixture was stirred at 0-5° C. for an additional 1 h, it wasallowed to warm to 25° C. and stirred at that temperature for 16 h. Thereaction was then poured slowly into an ice/water slurry and extractedwith dichloromethane (3×50 mL). The combined organic layers were driedover sodium sulfate and evaporated under reduced pressure to give ayellow solid. The product was purified by flash chromatography (MerckSilica gel 60, 230-400 mesh, 90/10 hexanes/ethyl acetate) to provide(3,4-dichloro-phenyl)-oxo-acetic acid methyl ester (1.59 g, 10% yield)as a yellow solid: EI-HRMS m/e calcd for C₉H₆O₃Cl₂ (M⁺) 231.9694, found231.9698.

To a dry round bottom flask, fitted with a Dean Stark: trap filled with3 Å molecular sieves and a reflux condenser, under argon was placed(3,4-dichloro-phenyl)-oxo-acetic acid methyl ester (1.00 g, 4.29 mmol))and p-toluenesulfonylhydrazide (1.03 g, 4.29 mmol) in toluene (20 mL).The reaction was heated at 110° C. for 16 h, then was cooled to 25° C.and the solvent removed in vacuo to yield a light yellow solid. Theproduct was crystallized from hot methanol to afford(3,4-dichloro-phenyl)-(4-toluenesulfonylhydrazono)-acetic acid methylester (1.45 g, 84% yield) as an off white solid: EI-HRMS m/e calcdC₁₆H₁₄Cl₂N₂O₄S (M⁺) 400.0051, found 400.0057.

In a dry flask under argon was placed a solution of(3,4-dichloro-phenyl)-(4-toluenesulfonylhydrazono)-acetic acid methylester (1.45 g, 3.61 mmol) in dichloromethane (20 mL) containingtriethylamine (0.55 mL, 3.97 mmol) at 25° C. The bright yellow solutionwas then stirred at 25° C. for 1 h, then the solvent was removed invacuo to yield a bright yellow solid. The product was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 7/1/0.5hexanes/dichloromethane/methanol) to furnishdiazo-(3,4-dichloro-phenyl)-acetic acid methyl ester (814 mg, 92% yield)as a bright yellowish orange solid: EI-HRMS m/e calcd for C₉H₆Cl₂N₂O₂(M⁺) 243.9806, found 243.9800.

In a dry flask under argon was placed diazo-(3,4-dichloro-phenyl)-aceticacid methyl ester (350 mg 1.4 mmol) to which was added dichloromethane(10 mL) and cyclopentanol (0.25 mL, 2.8 mmol). The solution was stirredat 25° C. and as rhodium (II) acetate dimer (13 mg, 0.028 mmol) wasadded, the immediate evolution of gas was noted and the color changedfrom bright yellow to an aquagreen color. After the solution was stirredat 25° C. for 1 h, it was then poured into water and the layers wereseparated. The aqueous layer was washed with dichloromethane (3×15 mL)and the organic layers were then combined, dried over sodium sulfate andconcentrated in vacuo. The residual material was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 95/5 hexanes/ethylacetate) to give rac-cyclopentyloxy-(3,4-dichloro-phenyl)-acetic acidmethyl ester (273 mg, 64% yield) as a clear colorless oil: EI-HRMS m/ecalcd for C₁₄H₁₆Cl₂O₃ (M⁺) 302.0477, found 302.0484.

A solution of rac-cyclopentyloxy-(3,4-dichloro-phenyl)-acetic acidmethyl ester (266 mg, 0.877 mmol) in ethanol (10 mL) was treated with asolution of potassium hydroxide (123 mg, 2.19 mmol) in water (1 mL) andthe mixture was stirred at 25° C. After 3 h, the reaction was dilutedwith water (5 mL) and the ethanol was removed in vacuo. The aqueouslayer was then acidified to pH 2 with 1 N hydrochloric acid andextracted with dichloromethane (3×15 mL). The combined organic layerswere dried over sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by flash chromatography (Merck Silicagel 60, 230-400 mesh, 95/5 chloroform/methanol plus 1% acetic acid) toafford rac-cyclopentyloxy-(3,4-dichloro-phenyl)-acetic acid (223 mg, 88%yield) as a white solid, mp 87.5-89.9 IC; EI-HRMS m/e calcd forC₁₃H₁₄Cl₂O₃ (M⁺) 288.0320, found 288.0332.

A solution of rac-cyclopentyloxy-(3,4-dichloro-phenyl)-acetic acid (52mg, 0.17 mmol) in dichloromethane (10 mL) was treated withO—(1H-benzotriazolo-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(HBTU) (72 mg, 0.19 mmol), diisopropyl-ethylamine (0.09 mL, 0.52 mmol)and 2-aminothiazole (26 mg, 0.25 mmol). The resulting brownish-orangesolution was then stirred 16 h at 25° C. The reaction was then dilutedwith water (10 mL) and extracted with ethyl acetate (3×15 mL). Thecombined organic layers were washed with water (1×10 mL), 1N sodiumhydroxide solution (1×10 mL), 1N hydrochloric acid (1×10 mL) and brine(1×10 mL), then were dried over sodium sulfate and concentrated invacuo. The product was purified by flash chromatography (Merck Silicagel 60, 230-400 mesh, 90/10 hexanes/ethyl acetate) to furnishrac-2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(45 mg, 70% yield) as a white foam: EI-HRMS m/e calcd for C₁₆H₁₆Cl₂O₂N₂S(M⁺) 370.0309, found 370.0309.

Example 2 Preparation ofrac-2-cyclohexyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide

In a dry 25 mL round bottom flask under argon was placeddiazo-(3,4-dichloro-phenyl)-acetic acid methyl ester (from Example 1,550 mg 2.24 mmol) and cyclohexanol (0.47 mL, 4.49 mmol) indichloromethane (10 mL). The solution was stirred at 25° C. and asrhodium (II) acetate dimer (20 mg, 0.045 mmol) was added, the immediateevolution of gas was observed and the color changed from bright yellowto an aquagreen color. After the solution was stirred at 25° C. for 1 h,it was poured into water and the layers were separated. The aqueouslayer was washed with dichloromethane (3×15 mL) and the combined organiclayers were dried over sodium sulfate and evaporated under reducedpressure. The residual oil was purified by flash chromatography (MerckSilica gel 60, 230-400 mesh, 98/2 hexanes/ethyl acetate) to furnishrac-cyclohexyloxy-(3,4-dichloro-phenyl)-acetic acid methyl ester (527mg, 74% yield) as a clear colorless oil: EI-HRMS m/e calcd forC₁₅H₁₈Cl₂O₃ (M⁺) 316.0633, found 316.0646.

A solution of rac-cyclohexyloxy-(3,4-dichloro-phenyl)-acetic acid methylester (527 mg, 1.66 mmol) in ethanol (15 mL) was treated with a solutionof potassium hydroxide (233 mg, 4.15 mmol) in water (2 mL) and themixture was stirred at 25° C. After 3 h, the reaction was diluted withwater (5 mL), and the ethanol was removed in vacuo. The aqueous layerwas then acidified to pH 2 with 1N hydrochloric acid and extracted withdichloromethane (3×15 mL). The combined organic layers were dried oversodium sulfate, filtered and concentrated in vacuo. The residual oil waspurified by flash chromatography (Merck Silica gel 60, 230-400 mesh,95/5 chloroform/methanol plus 1% acetic acid) to giverac-cyclohexyloxy-(3,4-dichloro-phenyl)-acetic acid (487 mg, 97% yield)as a colorless oil: EI-HRMS m/e calcd for C₁₄H₁₆Cl₂O₃ (M⁺) 302.0477,found 302.0486.

A solution of rac-cyclohexyloxy-(3,4-dichloro-phenyl)-acetic acid (102mg, 0.34 mmol) in dichloromethane (10 mL) was treated withbenzotriazol-1-yloxy-(dimethylamino)phosphonium hexafluorophosphate(BOP) reagent (223 mg, 0.51 mmol), triethylamine (0.14 mL, 0.52 mmol),and 2-aminothiazole (51 mg, 0.51 mmol) at 25° C. After the resultingbrownish-orange solution was stirred 16 h at 25° C., it was diluted withwater (10 mL) and extracted with ethyl acetate (3×15 mL). The combinedorganic layers were washed with water (1×10 mL), 1N sodium hydroxidesolution (1×10 mL), 1N hydrochloric acid (1×10 mL), and brine (1×10 mL),then were dried over sodium sulfate and evaporated in vacuo. The productwas purified by flash chromatography (Merck Silica gel 60, 230-400 mesh,90/10 hexanes/ethyl acetate) to furnishrac-2-cyclohexyloxy-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(115 mg, 88% yield) as a white foam: EI-HRMS m/e calcd forC₁₇H₁₈Cl₂O₂N₂S (M⁺) 384.0466, found 384.0469.

Example 3 Preparation ofrac-2-(cyclohex-2-enyloxy)-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide

In a dry 25 mL round bottom flask under argon was placeddiazo-(3,4-dichloro-phenyl)-acetic acid methyl ester (from Example 1,552 mg, 2.25 mmol), dichloromethane (10 mL) and rac-2-cyclohexen-1-ol(0.45 mL, 4.51 mmol). The solution was stirred at 25° C. and then therhodium (II) acetate dimer (20 mg, 0.045 mmol) was added. Gas evolutionbegan immediately and the color changed from bright yellow to anaquagreen color. After the solution was stirred at 25° C. for a periodof 1 h, it was poured into water and the layers were separated. Theaqueous layer was washed with dichloromethane (3×15 mL), then thecombined organic layers were dried over sodium sulfate and evaporatedunder reduced pressure. The residual oil was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 98/2 hexanes/ethylacetate to afford rac-(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-aceticacid methyl ester (552 mg, 78% yield) as a, light yellow oil: EI-HRMSm/e calcd for C₁₅H₁₆Cl₂O)₃ (M⁺) 314.0468, found 314.0476.

A solution of rac-(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetic acidmethyl ester (552 mg, 0.877 mmol) in ethanol (10 mL) to was treated witha solution of potassium hydroxide (246 mg, 4.37 mmol) and water (2 mL)and the mixture was stirred at 25° C. After 3 h, the reaction wasdiluted with water (10 mL) and the ethanol was removed in vacuo. Theaqueous layer was then acidified to pH 2 with 1N hydrochloric acid andextracted with dichloromethane (3×15 mL). The combined organic layerswere dried over sodium sulfate, filtered and concentrated in vacuo. Theproduct was purified by flash chromatography (Merck Silica gel 60,230-400 mesh, 95/5 chloroform/methanol plus 1% acetic acid) to giverac-(cyclohex-2-eyloxy)-(3,4-dichloro-phenyl)-acetic acid (520 mg, 99%yield) as a yellow oil: EI-HRMS m/e calcd for C₁₄H₁₄Cl₂O₃ (M⁺) 300.0320,found 300.0324.

A solution of rac-(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetic acid(89 mg, 0.28 mmol) in dichloromethane (10 mL) was treated with BOPreagent (187 mg, 0.42 mmol), triethylamine (0.12 mL, 0.85 mmol), and2-aminothiazole (42 mg, 0.42 mmol) at 25° C. The resultingbrownish-orange solution was then stirred 16 h at 25° C., then wasdiluted with water (10 mL) and extracted with ethyl acetate (3×15 mL).The combined organic layers were washed with water (1×10 mL), 1N sodiumhydroxide solution (1×10 mL), 1N hydrochloric acid (1×10 mL), and brine(1×10 mL), then were dried over sodium sulfate and evaporated in vacuo.The residue was purified by flash chromatography (Merck Silica gel 60,230-400 mesh, 95/5 hexanes/ethyl acetate) to providerac-(cyclohex-2-enyloxy)-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(99 mg, 92% yield) as a white foam: EI-HRMS m/e calcd for C₁₇H₁₆Cl₂O₂N₂S(M⁺) 382.0309, found 382.0308.

Example 4 Preparation ofrac-2-(3,4-dichloro-phenyl)-2-[(tetrahydro-pyran-4-yl)oxy]-N-thiazol-2-yl-acetamide

In a dry 25 mL round bottom flask under argon was placeddiazo-(3,4-dichloro-phenyl)-acetic acid methyl ester (from Example 1,614 mg, 2.51 mmol), dichloromethane (10 mL) and tetrahydro-4H-pyran-4-ol(0.50 mL, 5.01 mmol). The solution was stirred at 25° C. and thenrhodium (II) acetate dimer (22 mg, 0.05 mmol) was added. Gas evolutionbegan immediately and the color changed from bright yellow to anaquagreen color. After the solution was stirred at 25° C. for 1 h, itwas poured into water (10 mL) and the layers were separated. The aqueouslayer was washed with dichloromethane (3×15 mL) and the combined organiclayers were dried over sodium sulfate and in vacuo. The residualmaterial was purified by flash chromatography (Merck Silica gel 60,230-400 mesh, 98/2 hexanes/ethyl acetate) to furnishrac-(3,4-dichloro-phenyl)-[(tetrahydro-pyran-4-yl)oxy]-acetic acidmethyl ester (598 mg, 75% yield) as a clear colorless oil: EI-HRMS m/ecalcd for C₁₄H₁₆Cl₂O₄ (M⁺) 318.0426, found 318.0412.

A solution ofrac-(3,4-dichloro-phenyl)-[(tetrahydro-pyran-4-yl)oxy]-acetic acidmethyl ester (598 mg, 1.87 mmol) in ethanol (15 mL) was treated with asolution of potassium hydroxide (262 mg, 4.68 mmol) and water (2 mL) andthe mixture was allowed to stir at 25° C. After 3 h, the reaction wasdiluted with water (10 mL) and the ethanol was removed in vacuo. Theaqueous layer was then acidified to pH 2 with 1N hydrochloric acid andextracted with dichloromethane (3×15 mL). The combined organic layerswere dried over sodium sulfate, filtered and evaporated under reducedpressure. The reaction product was purified by flash chromatography(Merck Silica gel 60, 230-400 mesh, 95/5 chloroform/methanol plus 1%acetic acid) to affordrac-(3,4-dichloro-phenyl)-[(tetrahydro-pyran-4-yl)oxy]-acetic acid (544mg, 95% yield) as a clear colorless oil: EI-HRMS m/e calcd forC₁₃H₁₄Cl₂O₄ (M⁺) 304.0269, found 304.0259.

A solution ofrac-(3,4-dichloro-phenyl)-[(tetrahydro-pyran-4-yl)oxy]-acetic acid (90mg, 0.30 mmol) in dichloromethane (10 mL) was treated with BOP reagent(195 mg, 0.44 mmol), triethylamine (0.12 mL, 0.88 mmol), and2-aminothiazole (44 mg, 0.44 mmol) at 25° C. After the resultingbrownish-orange solution was stirred 16 h at 25° C., it was diluted withwater (10 ml) and extracted with ethyl acetate (3×15 mL). The combinedorganic layers were washed with water (1×10 mL), 1N sodium hydroxidesolution (1×10 mL), 1N hydrochloric acid (1×10 mL) and brine (1×10 mL),then were dried over sodium sulfate and evaporated in vacuo. Theresidual material was purified by flash chromatography (Merck Silica gel60, 230-400 mesh, 90/10 hexanes/ethyl acetate) to giverac-2-(3,4-dichloro-phenyl)-2-[(tetrahydro-pyran-4-yl)oxy]-N-thiazol-2-yl-acetamide(98 mg, 86% yield) as a white foam: EI-HRMS m/e calcd for C₁₆H₁₆Cl₂O₃N₂S(M⁺) 386.0258, found 386.0261.

Example 5 Preparation ofrac-2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-pyridin-2-yl-acetamide

A solution of rac-cyclopentyloxy-(3,4-dichloro-phenyl)-acetic acid (fromExample 1, 50 mg, 0.17 mmol) and triethylamine (0.07 mL, 0.52 mmol) intoluene (5 mL), previously cooled to 0° C. was treated with2,4,6-trichlorobenzoyl chloride (0.03 mL, 0.19 mmol) and the mixture wasstirred at 0° C. After 1 h, 2-aminopyridine (20 mg, 0.21 mmol) and4-dimethylaminopyridine (5 mg, 0.035 mmol) were added and the stirringwas continued for 1 h at 0° C. The reaction was checked for completion,then it was diluted with water (10 mL) and extracted withdichloromethane (3×10 mL). The combined organic extracts were dried oversodium sulfate, filtered and concentrated in vacuo. The product waspurified by chromatography (Biotage Flash 12M column, 80/20hexanes/ethyl acetate) to giverac-2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-N-pyridin-2-yl-acetamide(48 mg, 76% yield) as a white foam: EI-HRMS m/e calculated forC₁₈H₁₈N₂O₂Cl₂ (M⁺) 364.0745, found 364.0746.

Example 6 Preparation ofrac-2-(3-chloro-4-methanesulfonyl-phenyl)-2-cyclopentyloxy-N-thiazol-2-yl-acetamide

A solution of aluminum chloride (105.3 g, 789.4 mmol) in chloroform (300mL), cooled to 0° C., was treated with methyl oxalyl chloride (46.52 mL,505.8 mmol) in chloroform (300 mL) and the reaction was stirred at 0° C.After 30 min, the reaction was treated with a solution of2-chlorothioanisole (75.00 g, 472.7 mmol) in chloroform (300 mL) and thestirred reaction was allowed to equilibrate to 25° C. After 4 h, thereaction mixture was poured slowly into ice (2 L) and allowed to sit for15 min. It was then filtered through celite to remove the aluminum saltsand the filtrate was extracted with dichloromethane (3×50 mL). Theorganic extracts were then washed with saturated sodium bicarbonate(1×1100 mL), dried over magnesium sulfate, filtered and evaporated underreduced pressure. The product,(3-chloro-4-methylsulfanyl-phenyl)-oxo-acetic acid methyl ester (39.22g, 34% yield), which needed no further purification was isolated as alight yellow solid, mp 67.9-70.2° C.; EI-HRMS m/e calcd for C₁₀H₉ClSO₃(M⁺) 243.9961, found 243.9958.

To a clear solution of (3-chloro-4-methylsulfanyl-phenyl)-oxo-aceticacid methyl ester (5.00 g, 20.43 mmol) in methanol (100 mL) and water(10 mL) at 25° C. was added oxone (37.68 g, 61.29 mmol) in one portionand pH 4 phosphate buffer (5 mL). After the reaction was stirred for 5h, it was concentrated in vacuo to remove methanol, then was dilutedwith water (50 mL) and was extracted with ethyl acetate (3×50 mL). Thecombined organic extracts were dried over magnesium sulfate, filteredand evaporated under reduced pressure. The product was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 70/30 hexanes/ethylacetate) to provide (3-chloro-4-methane-sulfonyl-phenyl)-oxo-acetic acidmethyl (3.67 g, 65% yield) as a light yellow solid, mp 101.7-121.2° C.;EI-HRMS m/e calcd for C₁₀H₉ClSO₅ (M⁺) 275.9859, found 275.9857.

A solution of (3-chloro-4-methanesulfonyl-phenyl)-oxo-acetic acid methylester (3.67 g, 13.26 mmol) and p-toluenesulfonylhydrazide (3.21 g, 17.24mmol) in toluene (50 mL) was refluxed for 16 h in a flask fitted aDean-Stark trap filled with 3 A molecular sieves. The reaction was thencooled to 25° C. and concentrated in vacuo. The residual material wasflash chromatographed (Merck Silica gel 60, 230-400 mesh, 70/30hexanes/ethyl acetate) to provide(3-chloro-4-methanesulfonyl-phenyl)-(4-toluene-sulfonylhydrazono)-aceticacid methyl ester (3.82 g, 65% yield) as an off white solid. Thecompound was used per se in the subsequent transformation.

A solution of(3-chloro-4-methanesulfonyl-phenyl)-(4-toluenesulfonyl-hydrazono)-aceticacid methyl ester (3.82 g, 8.5 mmol) and triethylamine (1.3 mL, 9.35mmol) in dichloromethane (40 mL) was stirred at 25° C. After 1 h, thereaction was evaporated under reduced pressure and the resulting residuewas purified by flash chromatography (Merck Silica gel 60, 230-400 mesh,60/40 hexanes/ethyl acetate) to give(3-chloro-4-methanesulfonyl-phenyl)-diazo-acetic acid methyl ester (978mg, 40% yield) as a bright yellowish orange solid, mp 102.7-106.5° C.;EI-HRMS m/e calcd for C₁₀H₉N₂ClSO₄ (M⁺) 287.9972, found 287.9979.

A solution of (3-chloro-4-methanesulfonyl-phenyl)-diazo-acetic acidmethyl ester (489 mg, 1.69 mmol) in dichloromethane (10 mL) at 25° C.was treated with cyclopentanol (0.38 mL, 4.23 mmol) followed by rhodium(II) acetate dimer (15 mg, 0.034 mmol). After the resulting solution wasstirred at 25° C. for 1 h, it was diluted with dichloromethane (10 mL),poured into water (15 mL) and extracted with dichloromethane (3×10 mL).The combined organic extracts were dried over sodium sulfate, filteredand concentrated in vacuo. The product was purified by chromatography(Biotage Flash 40S column, 75/25 hexanes/ethyl acetate) to affordrac-(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetic acidmethyl ester (395 mg, 67% yield) as a colorless oil: EI-HRMS m/e calcdfor C₁₅H₁₉ClSO₅ (M⁺) 346.0642, found 346.0643.

A solution ofrac-(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetic acidmethyl ester (395 mg, 1.14 mmol) in ethanol (15 mL) at 25° C. wastreated with a solution of potassium hydroxide (320 mg, 5.69 mmol) inwater (3 mL) and the mixture was stirred at 25° C. After 3 h, thereaction was diluted with water and evaporated under reduced pressure.The concentrate was acidified to pH 2 with an aqueous solution of 1Nhydrochloric acid and extracted with dichloromethane (3×15 mL). Thecombined organic extracts were dried over sodium sulfate, filtered andevaporated in vacuo. The residual oil was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 50/50 hexanes/ethylacetate plus 1% acetic acid) to furnishrac-(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetic acid (364mg, 96% yield) as a colorless oil: EI-HRMS m/e calcd for C₁₄H₁₇ClSO₅(M⁺) 332.0485, found 332.0486.

To a stirred solution ofrac-(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetic acid (50mg, 0.15 mmol) in dichloromethane (10 mL) at 25° C. was added2-aminothiazole (23 mg, 0.23 mmol), BOP reagent (100 mg, 0.23 mmol) andtriethylamine (0.06 mL, 0.45 mmol). The mixture was stirred at 25° C.for 16 h, then it was diluted with water (10 mL) and extracted withdichloromethane (3×20 mL). The combined organic layers were washed withwater (1×10 mL), 1N sodium hydroxide (1×10 mL), 1N hydrochloric acid(1×10 mL) and brine (1×10 mL), then were dried over sodium sulfate,filtered and concentrated in vacuo. The product was purified bychromatography (Biotage Flash 40S column, 60/40 hexanes/ethyl acetate)to giverac-2-(3-chloro-4-methanesulfonyl-phenyl)-2-cyclopentyloxy-N-thiazol-2-yl-acetamide(44mg, 71% yield) as a white solid: EI-HRMS m/e calculated forC₁₇H₁₉N₂O₄S₂Cl (M⁺) 414.0475, found 414.0481.

Example 7 Preparation ofrac-1-[(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetyl]-3-methyl-urea

A cooled (0° C.) solution ofrac-(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetic acid(Example 6; 100 mg, 0.30 mmol) in fluorobenzene (2.5 mL) andN,N-dimethylformamide (1.8 μL). was treated with a 2.0 M solution ofoxalyl chloride in dichloromethane (0.18 mL, 0.36 mmol). Immediately avigorous gas evolution was observed, and the mixture was stirred at 25°C. for 1 h and became light yellow in color. Methyl urea (97 mg, 0.90mmol) was then added and after the reaction was heated at 70° C. for 10min, pyridine (0.048 mL, 0.60 mmol) was added and the reaction wasmaintained at 70° C. for 1 h. The cooled mixture was diluted with ethylacetate (5 mL) then was filtered through Celite to remove insolublematerials and the filtrate concentrated in vacuo. The concentrate waswashed with 3N hydrochloric acid (1×20 mL), saturated sodium bicarbonate(1×15 mL) and brine (1×15 mL), then was dried over sodium sulfate,filtered and evaporated under reduced pressure. The product was purifiedby chromatography (Biotage Flash 40S column, 50/50 hexanes/ethylacetate) to providerac-1-[(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetyl]-3-methyl-urea(78 mg, 67% yield) as a white foam: FAB-HRMS m/e calculated forC₁₆H₂₁N₂O₅SCl (M+H)⁺ 389.0938, found 389.0943.

Example 8 Preparation ofrac-2-(3-chloro-4-methanesulfonyl-phenyl)-2-(cyclohex-2-enyloxy)-N-thiazol-2-yl-acetamide

A solution of (3-chloro-4-methanesulfonyl-phenyl)-diazo-acetic acidmethyl ester (Example 6; 489 mg, 1.69 mmol) in dichloromethane (10 ML)at 25° C. was treated with 2-cyclohexen-1-ol (0.42 mL, 4.23 mmol)followed by rhodium (II) acetate dimer (15 mg, 0.034 mmol) and theresulting solution was stirred at 25° C. for 1 h. The reaction mixturewas diluted with dichloromethane (10 mL), then was poured into water (15mL) and extracted with dichloromethane (3×10 mL). The combined organiclayers were dried over sodium sulfate, filtered and concentrated invacuo. The product was purified by chromatography (Biotage Flash 40Scolumn, 75/25 hexanes/ethyl acetate) providedrac-(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetic acidmethyl ester (350 mg, 58% yield) as a colorless oil: EI-HRMS m/e calcdfor C₁₆H₁₉ClSO₅ (M⁺) 358.0642, found 358.0640.

A solution ofrac-(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetic acidmethyl ester (350 mg, 0.98 mmol) in ethanol (15 mL) at 25° C. wastreated with a solution of potassium hydroxide (273 mg, 4.88 mmol) inwater (2.5 mL) and the solution was stirred at 25° C. After 3 h, thereaction was diluted with water and concentrated under reduced pressure.The concentrate was acidified to pH 2 with an aqueous solution of 1Nhydrochloric acid and extracted with dichloromethane (3×15 mL). Thecombined organic layers were dried over sodium sulfate, filtered andevaporated under reduced pressure. The residual material was purified byflash chromatography (Merck Silica gel 60, 230-400 mesh, 50/50hexanes/ethyl acetate plus 1% acetic acid) to giverac-(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetic acid(265 mg, 79% yield) as a colorless oil: EI-HRMS m/e calcd forC₁₅H₁₇ClSO₅ (M⁺) 344.0485, found 344.0494.

To a solution ofrac-(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetic acid(50 mg, 0.15 mmol) in dichloromethane (10 mL) at 25° C. was added2-aminothiazole (22 mg, 0.22 mmol), BOP reagent (96.2 mg, 0.22 mmol) andtriethylamine (0.06 mL, 0.44 mmol). The mixture was stirred at 25° C.for 16 h, then was diluted with water (10 mL) and extracted withdichloromethane (3×20 mL). The combined organic layers were washed withwater (1×10 mL), 1N sodium hydroxide (1×10 mL), 1N hydrochloric acid(1×10 mL) and brine (1×10 mL), then were dried over sodium sulfate,filtered and evaporated under reduced pressure. The product was purifiedby chromatography (Biotage Flash 40S column, 60/40 hexanes/ethylacetate) to furnishrac-2-(3-chloro-4-methanesulfonyl-phenyl)-2-(cyclohex-2-enyloxy)-N-thiazol-2-yl-acetamide(39 mg, 63% yield) as a glassy solid: EI-HRMS m/e calculated forC₁₈H₁₉N₂O₄S₂Cl (M⁺) 426.0475, found 426.0479.

Example 9 Preparation ofrac-1-[(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetyl]-3-methyl-urea

A cooled (0° C.) mixture ofrac-(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetic acid(from Example 8100 mg, 0.29 mmol), fluorobenzene (2.5 mL) andN,N-dimethylformamide (1.8 μL). was treated with a 2.0 M solution ofoxalyl chloride in dichloromethane (0.18 mL, 0.36 mmol) which caused avigorous evolution of gas. The reaction was then stirred at 25° C. for 1h and became light yellow in color. After methyl urea (64 mg, 0.87 mmol)was added, the reaction was heated at 70° C. for 10 min, then pyridine(0.048 mL, 0.60 mmol) was added and the mixture was maintained at 70° C.for 1 h. The cooled reaction was diluted with ethyl acetate (5 mL), thenwas filtered through celite to remove insoluble materials and thefiltrate was concentrated in vacuo. The concentrate was washed with 3Nhydrochloric acid (1×20 mL), saturated sodium bicarbonate (1×15 mL), andbrine (1×15 mL), then was dried over sodium sulfate, filtered andevaporated under reduced pressure. The reaction product was purified bychromatography (Biotage Flash 40S column, 50/50 hexanes/ethyl acetate)to give1-[(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetyl]-3-methyl-urea(63 mg, 54% yield) as a white foam: FAB-HRMS m/e calculated forC₁₇H₂₁N₂O₅SCl (M+H)⁺ 401.0938, found 401.0921.

Example 10 Preparation ofrac-2-cyclopentylsulfanyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide

A solution of diazo-(3,4-dichloro-phenyl)-acetic acid methyl ester(Example 1; 193 mg, 0.79 mmol) in dichloromethane (10 mL) at 25° C. wastreated with cyclopentyl mercaptan (0.21 mL, 1.97 mmol) followed byrhodium (II) acetate dimer (9 mg, 0.020 mmol) and the solution wasstirred at 25° C. for 1 h. During this time no evolution of gas wasdetected, and examination of the black solution by thin layerchromatography indicated that only starting material was present. Thereaction was heated to reflux and a second portion of rhodium (II)acetate dimer (10 mg, 0.024 mmol) was added and as the mixture wasstirred at reflux for 110 min, a vigorous evolution of gas was observed.The reaction mixture was diluted with dichloromethane (10 mL), then waspoured into water (15 mL) and extracted with dichloromethane (3×10 mL).The combined organic extracts were dried over sodium sulfate, filteredand evaporated under reduced pressure. The residual oil was purified byflash chromatography (Merck Silica gel 60, 230-400 mesh, 95/5hexanes/ethyl acetate) to furnishrac-cyclopentylsulfanyl-(3,4-dichloro-phenyl)-acetic acid methyl ester(148 mg, 59% yield) as a colorless oil: EI-HRMS m/e calcd forC₁₄H₁₆Cl₂SO₂(M⁺) 318.0248, found 318.0244.

A solution of rac-cyclopentylsulfanyl-(3,4-dichloro-phenyl)-acetic acidmethyl ester (50 mg, 0.16 mmol) in ethanol (3 mL) at 25° C. was treatedwith a solution of potassium hydroxide (44 mg, 0.79 mmol) in water (1mL) and the mixture was stirred at 25° C. After 3 h, the reaction wasdiluted with water and evaporated under reduced pressure. Theconcentrate was acidified to pH 2 with art aqueous solution of 1Nhydrochloric acid and extracted with dichloromethane (3×15 mL). Thecombined organic extracts were dried over sodium sulfate, filtered andconcentrated in vacuo. The residual material was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 50/50 hexanes/ethylacetate plus 1% acetic acid) to affordrac-cyclopentylsulfanyl-(3,4-dichloro-phenyl)-acetic acid (43 mg, 90%yield) as a colorless oil: EI-HRMS m/e calcd for C₁₃H₁₄Cl₂SO₂ (M⁺)304.0091, found 304.0101.

Cyclopentylsulfanyl-(3,4-dichloro-phenyl)-acetic (43 mg, 0.14 mmol) wasdissolved in dichloromethane (10 mL) and to this solution at 25° C. wasadded 2-aminothiazole (21 mg, 0.21 mmol), BOP reagent (92 mg, 0.21 mmol)and triethylamine (0.06 mL, 0.42 mmol). The reaction mixture was stirredat 25° C. for 16 h, then was diluted with water (10 mL) and extractedwith dichloromethane (3×15 mL). The combined organic layers were washedwith water (1×10 mL), 1N sodium hydroxide (1×10 mL), 1N hydrochloricacid (1×10 mL) and brine (1×10 mL), then were dried over sodium sulfate,filtered and evaporated under reduced pressure. The product was purifiedby chromatography (Biotage Flash 12M column, 80/20 hexanes/ethylacetate) to furnishrac-2-cyclopentylsulfanyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(40 mg, 74% yield) as a colorless oil: EI-HRMS m/e calculated forC₁₆H₁₆N₂OS₂Cl₂ (M⁺) 386.0081, found 386.0080.

Example 11 Preparation ofrac-2-cyclopentanesulfonyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide

To a solution ofrac-2-cyclopentylsulfanyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(Example 10; 34 mg, 0.088 mmol) in methanol (2 mL) was added a solutionof sodium periodate (34 mg, 0.16 mmol) in water (1 mL) and the mixturewas stirred at 25° C. After 6 h, the precipitate was filtered off andwashed with dichloromethane (15 mL). The organic layer was set aside andthe aqueous layer was extracted with dichloromethane (3×10 mL). Thecombined organic layers were dried over sodium sulfate, filtered andevaporated under reduced pressure. Chromatography of the residue(Biotage Flash 12M column, 50/50 hexanes/ethyl acetate) affordedrac-2-cyclopentanesulfinyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(23 mg, 66% yield) as a colorless oil: EI-HRMS m/e calculated forC₁₆H₁₆N₂O₂S₂Cl₂ (M⁺) 402.0030, found 402.0035.

A solution ofrac-2-cyclopentanesulfinyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(20 mg, 0.05 mmol) in methanol (2 mL) was stirred at 25° C. as asolution of potassium permanganate (9 mg, 0.06 mmol) in water (0.5 mL)was added. The mixture was stirred at 25° C. for 30 min and then wasfiltered. The filter cake was washed with dichloromethane and thecombined filtrates were washed with sodium bicarbonate solution (10 mL)and brine (10 mL), then were dried over sodium sulfate, filtered andconcentrated in vacuo. The product was purified by chromatography(Biotage Flash 12M column, 50/50 hexanes/ethyl acetate) to providerac-2-cyclopentanesulfonyl-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(10mg, 48% yield) as a colorless oil: EI-HRMS m/e calculated forC₁₆H₁₆N₂O₃S₂Cl₂ (M⁺) 417.9979, found 417.9977.

Example 12 Preparation ofrac-1-[cyclopentyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea

A cooled (0° C.) solution ofrac-cyclopentyloxy-(3,4-dichloro-phenyl)-acetic acid (Example 1; 164 mg,0.57 mmol) in dichloromethane (10 mL) and N,N-dimethylformamide (onedrop) was treated with oxalyl chloride (2.0 M solution indichloromethane, 0.43 mL, 0.86 mmol). The reaction was stirred at 0° C.for 1 h, then 1,1,1,3,3,3-hexamethyldisilazane (0.42 mL, 2.0 mmol) wasadded and the resulting cloudy mixture was stirred at 25° C. for 16 h.The reaction was quenched with methanol (10 mL), washed with an aqueoussolution of 5% sulfuric acid (2×15 mL) and extracted withdichloromethane (3×10 mL). The combined organic extracts were washedwith brine (1×10 mL), then were dried over magnesium sulfate, filteredand concentrated in vacuo. The residue was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 60/40 hexanes/ethylacetate) to give rac-2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-acetamide(116 mg, 71% yield) as a white solid, mp 88.3-91.4° C.; FAB-HRMS m/ecalcd for C₁₃H₁₅NCl₂O₂ (M⁺) 288.0558, found 288.0572.

A solution of rac-2-cyclopentyloxy-2-(3,4-dichloro-phenyl)-acetamide(116 mg, 0.40 mmol) in toluene (5 mL) was treated with methyl isocyanate(0.04 mL, 0.60 mmol). The resulting solution was heated to reflux for 24h, then was cooled and was evaporated under reduced pressure. Theresulting oil was purified by flash chromatography (Merck Silica gel 60,230-400 mesh, 60/40 hexanes/ethyl acetate) to furnish1-[cyclopentyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea (30 mg,41% yield) as a colorless oil: EI-HRMS m/e calcd for C₁₅H₁₈N₂Cl₂O₃ (M⁺)288.0558, found 288.0572.

Example 13 Preparation ofrac-1-[(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea

A solution of rac-(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetic acid(Example 3; 409 mg, 1.36 mmol) in dichloromethane (10 mL) andN,N-dimethylformamide (one drop) cooled to 0° C. was treated with oxalylchloride (2.0 M solution in dichloromethane, 0.95 mL, 1.90 mmol). Thereaction was stirred at 0° C. for 1 h, then1,1,1,3,3,3-hexamethyldisilazane (1.0 mL, 4.75 mmol) was added and theresulting cloudy mixture was stirred at 25° C. for 16 h. The reactionwas quenched with methanol (10 mL), washed with an aqueous solution of5% sulfuric acid (2×15 mL) and extracted with dichloromethane (3×10 mL).The combined organic extracts were washed with brine (1×10 mL), driedover magnesium sulfate, filtered and concentrated in vacuo. The reactionproduct was purified by flash chromatography (Merck Silica gel 60,230-400 mesh, 70/30 hexanes/ethyl acetate) to giverac-2-(cyclohex-2-enyloxy)-2-(3,4-dichloro-phenyl)-acetamide (311 mg,76% yield) as a white solid: 103.6-108.9° C.; EI-HRMS m/e calcd forC₁₄H₁₅NCl₂O₂(M⁺) 299.0479, found 299.0492.

A solution ofrac-2-(cyclohex-2-enyloxy)-2-(3,4-dichloro-phenyl)-acetamide (311 mg,1.04 mmol) in toluene (10 mL) was treated with methyl isocyanate (0.09mL, 1.55 mmol). The resulting solution was heated at reflux for 24 h andthen was concentrated in vacuo. The residual material was purified byflash chromatography (Merck Silica gel 60, 230-400 mesh, 90/10hexanes/ethyl acetate) to furnish1-[(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetyl] h-3-methyl-urea(238 mg, 64% yield) as a colorless oil: EI-HRMS m/e calcd forC₁₆H₁₈N₂Cl₂O₃ (M⁺) 356.0694, found 356.0694.

Example 14 Preparation ofrac-1-[cyclohexyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea

A solution of rac-cyclohexyloxy-(3,4-dichloro-phenyl)-acetic acid(Example 2; 364 mg, 1.20 mmol) in dichloromethane (10 mL) andN,N-dimethylformamide (one drop) cooled to 0° C. was treated with oxalylchloride (2.0M solution in dichloromethane, 0.84 mL, 1.68 mmol). Thereaction was stirred at 0° C. for 1 h, then1,1,1,3,3,3-hexamethyldisilazane (0.90 mL, 4.20 mmol) was added and thecloudy mixture was stirred at 25° C. for 16 h. At this time, thereaction was quenched with methanol (10 mL), washed with an aqueoussolution of 5% sulfuric acid (2×15 mL) and extracted withdichloromethane (3×10 mL). The combined organic extracts were washedwith brine (1×10 mL), then were dried over magnesium sulfate, filteredand evaporated under reduced pressure. The product was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 90/10 hexanes/ethylacetate) to afford rac-2-cyclohexyloxy-2-(3,4-dichloro-phenyl)-acetamide(311 mg, 76% yield) as a colorless oil: FAB-HRMS m/e calcd forC₁₄H₁₇NCl₂O₂ (M+H)⁺ 302.0714, found 302.0728.

A solution of rac-2-cyclohexyloxy-2-(3,4-dichloro-phenyl)-acetamide (291mg, 0.96 mmol) in toluene (10 mL) was treated with methyl isocyanate(0.09 mL, 1.44 mmol). The resulting solution was heated at reflux for 24h and then the cooled reaction was concentrated in vacuo. The reactionproduct was purified by flash chromatography (Merck Silica gel 60,230-400 mesh, 90/10 hexanes/ethyl acetate) to give1-[cyclohexyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea (301 mg,87% yield) as a colorless oil: EI-HRMS m/e calcd for C₁₆H₂₀N₂Cl₂O₃(M+H)⁺ 359.0929, found 359.0922.

Example 15 Preparation ofrac-[1-[(3,4-dichloro-phenyl)-(tetrahydro)-pyran-4-yloxy)-acetyl]-3-methyl-urea

A cooled (0° C.) solution ofrac-(3,4-dichloro-phenyl)-[(tetrahydro-pyran-4-yloxy)]-acetic acid(Example 4; 441 mg, 1.45 mmol) in dichloromethane (10 mL) andN,N-dimethylformamide (one drop) was treated with oxalyl chloride (2.0 Msolution in dichloromethane, 1.01 mL, 2.02 mmol). The reaction wasstirred at 0° C. for 1 h, then 1,1,1,3,3,3-hexamethyldisilazane (1.10mL, 5.06 mmol) was added and the resulting cloudy mixture was stirred at25° C. for 16 h. The reaction was quenched with methanol (10 mL), washedwith an aqueous solution of 5% sulfuric acid (2×15 mL) and extractedwith dichloromethane (3×10 mL). The combined organic extracts werewashed with brine (1×10 mL), then were dried over magnesium sulfate,filtered and evaporated under reduced pressure. The resulting residuewas purified by flash chromatography (Merck Silica gel 60, 230-400 mesh,70/30 hexanes/ethyl acetate) to providerac-2-(3,4-dichloro-phenyl)-2-(tetrahydro-pyran-4-yloxy)-acetamide (278mg, 63% yield) as a white solid, mp 81.9-83.6° C.; FAB-HRMS m/e calcdfor C₁₃H₁₅NCl₂O₃ (M+H)⁺ 303.0428, found 303.0426.

A solution ofrac-2-(3,4-dichloro-phenyl)-2-(tetrahydro-pyran-4-yloxy)-acetamide (278mg, 0.91 mmol) in toluene (10 mL) was treated with methyl isocyanate(0.08 mL, 1.37 mmol). The resulting solution was heated at reflux for 24h and then the cooled reaction was concentrated in vacuo. The resultingoil was purified by flash chromatography (Merck Silica gel 60, 230-400mesh, 20/80 hexanes/ethyl acetate) to afford[1[(3,4-dichloro-phenyl)-(tetrahydro-pyran-4-yloxy)-acetyl]-3-methyl-urea(70 mg, 21% yield) as a colorless oil: EI-HRMS m/e calcd forC₁₅H₁₈N₂Cl₂O₄ (M⁺) 360.0643, found 360.0865.

Example 16 Preparation ofrac-3-cyclopentyl-2-(3,4-dichloro-phenyl)-3-oxo-N-thiazol-2-yl-propionamide

To a solution of (3,4-dichloro-phenyl)-acetic acid (500 mg, 2.4 mmol)inN,N-dimethylformamide (15 mL) at 25° C. was added HBTU (1.02 g, 2.7mmol), 2-aminothiazole (360 mg, 3.6 mmol) and diisopropylethylamine(1.25 mL, 7.2 mmol). The reaction mixture was stirred for 16 h, then wasdiluted with water (10 mL) and extracted with ethyl acetate (3×15 mL).The combined organic layers were washed in turn with water (1×10 mL), 1Nsodium hydroxide (1×10 mL), 1 N hydrochloric acid (1×10 mL) and brine(1×10 mL), then were dried over sodium sulfate, filtered and evaporatedunder reduced pressure. The residual material was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 50150 hexanes/ethylacetate) to furnish rac-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(480 mg, 70% yield) as a light yellow solid, mp 169.8-172.3° C. EI-HRMSm/e calcd for C₁₁H₈N₂OSCl₂ (M⁺) 285.9734, found 285.9734.

To a solution of rac-2-(3,4-dichloro-phenyl)-N-thiazol-2-yl-acetamide(185 mg, 0.64 mmol) in tetrahydrofuran (15 mL), previously cooled to−78° C., was added slowly a 1.0 M solution of lithiumbis(trimethylsilyl)amide (1.3 mL, 1.3 mmol). The solution was stirredfor 15 min at −78° C., then cyclopentanecarbonyl chloride (0.08 mL, 0.64mmol) was dropwise added. The reaction mixture was stirred for anadditional 60 min at −78° C. before it was quenched by the addition of asaturated ammonium chloride solution (10 mL). The mixture was thenextracted with ethyl acetate (3×10 mL), dried over sodium sulfate,filtered and concentrated in vacuo. The product was purified by flashchromatography (Merck Silica gel 60, 230-400 mesh, 80/20 hexanes/ethylacetate) to afford3-cyclopentyl-2-(3,4-dichloro-phenyl)-3-oxo-N-thiazol-2-yl-propionamide(98 mg, 40% yield) as a yellow-orange solid: EI-HRMS m/e calcd forC₁₇H₁₆N₂O₂SCl₂ (M⁺) 382.0309, found 382.0309.

Biological Activity Examples

a) In Vitro Glucokinase Activity

Glucokinase Assay: Glucokinase (GK) was assayed by coupling theproduction of glucose-6-phosphate to the generation of NADH withglucose-6-phosphate dehydrogenase (G6PDH, 0.75-1 kunits/mg; BoehringerMannheim, Indianapolis, Ind.) from Leuconostoc mesenteroides as thecoupling enzyme (Scheme 2). Recombinant

Human liver GK1 was expressed in E. coli as a glutalhione S-transferasefusion protein (GST-GK) [Liang et al, 1995] and was purified bychromatography over a glutathione-Sepharose 4B affinity column using theprocedure provided by the manufacturer (Amersham Pharmacia Biotech,Piscataway, N.J.). Previous studies have demonstrated that the enzymaticproperties of native GK and GST-GK are essentially identical (Liang etal, 1995; Neet et al., 1990).

The assay was conducted at 25° C. in a flat bottom 96-well tissueculture plate from Costar (Cambridge, Mass.) with a final incubationvolume of 120 μL. The incubation mixture contained: 25 mM Hepes buffer(pH, 7.1), 25 mM KCl, 5 mM D-glucose, 1 mM ATP, 1.8 mM NAD, 2 mM MgCl₂,1 μM sorbitol-6-phosphate, 1 mM dithiothreitol, test drug or 10% DMSO,1.8 unit/ml G6PDH, and GK (see below). All organic reagents were >98%pure and were from Boehringer Mannheim with the exceptions of D-glucoseand Hepes that were from Sigma Chemical Co, St Louis, Mo. Test compoundswere dissolved in DMSO and were added to the incubation mixture minusGST-GK in a volume of 12 μl to yield a final DMSO concentration of 10%.This mix was preincubated in the temperature controlled chamber of aSPECTRAmax 250 microplate spectrophotometer (Molecular DevicesCorporation, Sunnyvale, Calif.) for 10 minutes to allow temperatureequilibrium and then the reaction was started by the addition of 20 μIGST-GK.

After addition of enzyme, the increase in optical density (OD) at 340 nmwas monitored over a 10 minute incubation period as a measure of GKactivity. Sufficient GST-GK was added to produce an increase in OD₃₄₀ of0.08 to 0.1 units over the 10 minute incubation period in wellscontaining 10% DMSO, but no test compound. Preliminary experimentsestablished that the GK reaction was linear over this period of timeeven in the presence of activators that produced a 5-fold increase in GKactivity. The GK activity in control wells was compared with theactivity in wells containing test GK activators, and the concentrationof activator that produced a 50% increase in the activity of GK, i.e.,the SC_(1.5), was calculated. All of the compounds of formula Idescribed in the Synthesis Examples had an SC_(1.5) less than or equalto 30 μM.

Liang, Y., Kesavan, P., Wang, L., Niswender, K., Tanizawa, Y., Permut,M. A., Magnuson, M., and Matschinsky, F. M. Variable effects ofmaturity-onset-diabetes-of-youth (MODY)-associated glucokinase mutationson the substrate interactions and stability of the enzyme. Biochem. J309: 167-173, 1995.

Neet, K., Keenan, R. P., and Tippett, P. S. Observation of a kineticslow transition in monomeric glucokinase. Biochemistry 29;770-777, 1990.

b) In vivo Activity

Glucokinase Activator in vivo Screen Protocol

C57BL/6J mice are orally dosed via gavage with Glucokinase (GK)activator at 50 mg/kg body weight following a two hour fasting period.Blood glucose determinations are made five times during the six hourpost-dose study period.

Mice (n=6) are weighed and fasted for a two hour period prior to oraltreatment. GK activators are formulated at 6.76 mg/ml in Gelucirevehicle (Ethanol:Gelucire44/14:PEG400q.s. 4:66:30 v/w/v. Mice are dosedorally with 7.5 μL formulation per gram of body weight to equal a 50mg/kg dose. Immediately prior to dosing, a pre dose (time zero) bloodglucose reading is acquired by snipping off a small portion of theanimals tail (˜1 mm) and collecting 15 μL blood into a heparinizedcapillary tube for analysis. Following GK activator administration,additional blood glucose readings are taken at 1, 2, 4, and 6 hours postdose from the same tail wound. Results are interpreted by comparing themean blood glucose values of six vehicle treated mice with six GKactivator treated mice over the six hour study duration. Compounds areconsidered active when they exhibit a statistically significant (p≦0.05)decrease in blood glucose compared to vehicle for two consecutive assaytime points.

Example A

Tablets containing the following ingredients can be produced in aconventional manner:

Ingredients mg per tablet Compound of formula (I) 10.0-100.0 Lactose125.0 Corn starch 75.0 Talc 4.0 Magnesium stearate 1.0

Example B

Capsules containing the following ingredients can be produced in aconventional manner:

Ingredients mg per capsule Compound of formula (I) 25.0 Lactose 150.0Corn starch 20.0 Talc 5.0

Upon reading the present specification, various alternative embodimentswill become obvious to the skilled artisan. These variations are to beconsidered within the scope and spirit of the subject invention which isonly to be limited by the claims that follow and their equivalents.

What is claimed is:
 1. A compound of the formula:

wherein R¹ and R² are independently hydrogen, halo, cyano, nitro, loweralkylthio, perfluoro loweralkylthio, lower alkyl sulfonyl, orperfluoro-lower alkyl sulfonyl; R³ is lower alkyl having from 2 to 4carbon atoms or a 5 to 7-membered ring which is cycloalkyl,cycloalkenyl, or heterocycloalkyl having one heteroatom selected fromoxygen and sulfur; R⁴ is —C(O)NHR⁵; R⁵ is hydrogen, lower alkyl, loweralkenyl, hydroxy lower alkyl, halo lower alkyl, —(CH₂₎ _(n)—C(O)—OR⁷,—C(O)—(CH₂)_(n)—C(O)—OR⁸; R⁷ and R⁸ are independently hydrogen or loweralkyl; X is oxygen, sulfur, sulfonyl or carbonyl; and the * indicates anasymmetric carbon atom; or a pharmaceutically acceptable salt thereof.2. The compound of claim 1, wherein said compound is of the formula:

wherein R¹ and R² are independently hydrogen, halo, lower alkylsulfonyl, or perfluoro-lower alkyl sulfonyl; R³ is a 5 to 7-memberedring which is cycloalkyl, cycloalkenyl, or heterocycloalkyl having oneheteroatom selected from oxygen and sulfur; R⁵ is lower alkyl; X isoxygen, sulfur, sulfonyl or carbonyl; and the * indicates an asymmetriccarbon atom, or a pharmaceutically acceptable salt thereof.
 3. Thecompound of claim 2, wherein R¹ and R² are independently halo or loweralkyl sulfonyl and R³ is a 5 to 7-membered ring which is cyclopentyl,cyclohexyl, cyclohexenyl, or a six-membered heterocycloalkyl having oneheteroatom selected from oxygen and sulfur.
 4. The compound of claim 3,wherein the heteroatom is oxygen.
 5. The compound of claim 4, wherein R⁵is methyl.
 6. The compound of claim 5, where X is oxygen.
 7. Thecompound of claim 6, wherein R¹ and R² are independently chloro ormethyl sulfonyl.
 8. The compound of claim 7, wherein R¹ and R² arechloro.
 9. The compound of claim 8 which is1-[cyclopentyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea.
 10. Thecompound of claim 8 which is1-[cyclohexyloxy-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea.
 11. Thecompound of claim 8 which is1-[(cyclohex-2-enyloxy)-(3,4-dichloro-phenyl)-acetyl]-3-methyl-urea. 12.The compound of claim 8 which is[1-[(3,4-dichloro-phenyl)-(tetrahydro-pyran-4-yloxy)-acetyl]-3-methyl-urea.13. The compound of claim 7, wherein R¹ is chloro and R² is methylsulfonyl.
 14. The compound of claim 13 which is1-[(3-chloro-4-methanesulfonyl-phenyl)-cyclopentyloxy-acetyl]-3-methyl-urea.15. The compound of claim 13 which is1-[(3-chloro-4-methanesulfonyl-phenyl)-(cyclohex-2-enyloxy)-acetyl]-3-methyl-urea.